Cell and Tissue Banking

, Volume 17, Issue 4, pp 745–756 | Cite as

The role of antioxidants in sperm freezing: a review

  • Fardin Amidi
  • Azar Pazhohan
  • Maryam Shabani NashtaeiEmail author
  • Mahshad Khodarahmian
  • Saeid Nekoonam
Full Length Review


Cryopreservation of spermatozoa is becoming more important because of new clinical requirements and current clinical practice. Despite the success of sperm cryopreservation this routinely used procedure induces serious detrimental changes in sperm function. Some researchers believe that cryopreservation is associated with DNA fragmentation and DNA single strand breaks in sperm. Mechanisms of cryodamage to human spermatozoa are thought to be multifactorial including: cold shock, osmotic stress, intracellular ice crystal formation, oxidative stress, and combinations of these conditions. Additives showing antioxidative properties reported to reduce the impact of ROS-induced and cold shock damages. Many studies exist as regards the effects of antioxidants on the cryopreservation aimed at improving the quality of post-thaw semen. Hence, this review will clarify results of recent applications of various antioxidants used in numerous research efforts to improve cryopreservation of spermatozoa. This review is to increase the understanding of the roles of these antioxidants concerning mechanisms which enhance resistance to cryodamage of spermatozoa.


Cryopreservation Sperm Cryodamage Oxidative stress 


  1. Agarwal A, Gupta S, Sikka S (2006a) The role of free radicals and antioxidants in reproduction. Curr Opin Obstet Gynecol 18:325–332PubMedCrossRefGoogle Scholar
  2. Agarwal A, Sharma RK, Nallella KP, Thomas AJ, Alvarez JG, Sikka SC (2006b) Reactive oxygen species as an independent marker of male factor infertility. Fertil Steril 86:878–885PubMedCrossRefGoogle Scholar
  3. Aitken RJ (1995) Free radicals, lipid peroxidation and sperm function. Reprod Fertil Dev 7(4):659–668PubMedCrossRefGoogle Scholar
  4. Akiyama T et al (1987) Genistein, a specific inhibitor of tyrosine-specific protein kinases. J Biol Chem 262:5592–5595PubMedGoogle Scholar
  5. Alvarez JG, Storey BT (1989) Role of glutathione peroxidase in protecting mammalian spermatozoa from loss of motility caused by spontaneous lipid peroxidation. Gamete Res 23:77–90PubMedCrossRefGoogle Scholar
  6. Alvarez JG, Touchstone JC, Blasco L, Storey BT (1987) Spontaneous lipid peroxidation and production of hydrogen peroxide and superoxide in human spermatozoa. Superoxide Dismutase as major enzyme protectant against oxygen toxicity. J Androl 8:338–348PubMedCrossRefGoogle Scholar
  7. Amidi F, Farshad A, Khor AK (2010) Effects of cholesterol-loaded cyclodextrin during freezing step of cryopreservation with TCGY extender containing bovine serum albumin on quality of goat spermatozoa. Cryobiology 61:94–99PubMedCrossRefGoogle Scholar
  8. AminiPour H, Tahmasbi AM, Naserian AA (2013) The influence of vitamin E on semen characteristics of ghezel rams in during cooling and frozen process. Eur J Zool Res 2(5):94–99Google Scholar
  9. Andreea A, Stela Z (2010) Role of antioxidant additives in the protection of the cryopreserved semen against free radicals. Rom Biotechnol Lett 15:33–41Google Scholar
  10. Anel-López L et al (2012) Reduced glutathione and Trolox (vitamin E) as extender supplements in cryopreservation of red deer epididymal spermatozoa. Anim Reprod Sci 135:37–46PubMedCrossRefGoogle Scholar
  11. Ashrafi I, Kohram H, Naijian H, Bahreini M, Poorhamdollah M (2011) Protective effect of melatonin on sperm motility parameters on liquid storage of ram semen at 5 C. Afr J Biotechnol 10:6670Google Scholar
  12. Askari H, Check J, Peymer N, Bollendorf A (1994) Effect of natural antioxidants tocopherol and ascorbic acids in maintenance of sperm activity during freeze-thaw process. Arch Androl 33:11–15PubMedCrossRefGoogle Scholar
  13. Asmus KD, Benasson RV, Bernier JL, Houssin R (1996) One-electron oxidation of ergothioneine and analogues investigated by pulse radiolysis: redox reaction involving ergothioneine and vitamin C. Biochem J 315:625–629PubMedPubMedCentralCrossRefGoogle Scholar
  14. Atmaca G (2004) Antioxidant effects of sulfur-containing amino acids. Yonsei Med J 45:776–788PubMedCrossRefGoogle Scholar
  15. Azawi OI, Hussein EK (2013) Effect of vitamins C or E supplementation to Tris diluent on the semen quality of Awassi rams preserved at 5 C. In: Veterinary Research Forum, vol 3. Faculty of Veterinary Medicine, Urmia University, Urmia, Iran, p 157Google Scholar
  16. Bailey JL, Lessard C, Jacques J, Brèque C, Dobrinski I, Zeng W, Galantino-Homer HL (2008) Cryopreservation of boar semen and its future importance to the industry. Theriogenology 70:1251–1259PubMedCrossRefGoogle Scholar
  17. Baiomy A, Mohamed A, Mottelib A, Briese A, Clauss M, Springorum A, Hartung J (2009) Effect of dietary selenium and vitamin E supplementation on productive and reproductive performance in rams. In: Sustainable animal husbandry: prevention is better than cure, vol 1. Proceedings of the 14th international congress of the international society for animal hygiene (ISAH), Vechta, Germany, 19th to 23rd July 2009. Tribun EU, pp 43–46Google Scholar
  18. Bajpai M, Doncel G (2003) Involvement of tyrosine kinase and cAMP-dependent kinase cross-talk in the regulation of human sperm motility. Reproduction 126:183–195PubMedCrossRefGoogle Scholar
  19. Ball BA (2008) Oxidative stress, osmotic stress and apoptosis: impacts on sperm function and preservation in the horse. Anim Reprod Sci 107:257–267PubMedCrossRefGoogle Scholar
  20. Beconi M, Francia C, Mora N, Affranchino M (1993) Effect of natural antioxidants on frozen bovine semen preservation. Theriogenology 40:841–851PubMedCrossRefGoogle Scholar
  21. Ben Abdallah F, Dammak I, Attia H, Hentati B, Ammar-Keskes L (2009) Lipid peroxidation and antioxidant enzyme activities in infertile men: correlation with semen parameter. J Clin Lab Anal 23:99–104PubMedCrossRefGoogle Scholar
  22. Ben Abdallah F, Zribi N, Ammar-Keskes L (2011) Antioxidative potential of Quercetin against hydrogen peroxide induced oxidative stress in spermatozoa in vitro. Andrologia 43:261–265PubMedCrossRefGoogle Scholar
  23. Bilodeau JF, Chatterjee S, Sirard MA, Gagnon C (2000) Levels of antioxidant defenses are decreased in bovine spermatozoa after a cycle of freezing and thawing. Mol Reprod Dev 55:282–288PubMedCrossRefGoogle Scholar
  24. Bilodeau J-F, Blanchette S, Gagnon C, Sirard M-A (2001) Thiols prevent H 2 O 2-mediated loss of sperm motility in cryopreserved bull semen. Theriogenology 56:275–286PubMedCrossRefGoogle Scholar
  25. Blesbois E, Caffin J (1992) ‘Serum like’ albumin of fowl seminal plasma and effects of albumin on fowl spermatozoa stored at 4 °C. Br Poult Sci 33:663–670PubMedCrossRefGoogle Scholar
  26. Bollwein H, Fuchs I, Koess C (2008) Interrelationship between plasma membrane integrity, mitochondrial membrane potential and DNA fragmentation in cryopreserved bovine spermatozoa. Reprod Domest Anim 43:189–195PubMedCrossRefGoogle Scholar
  27. Branco CS, Garcez ME, Pasqualotto FF, Erdtman B, Salvador M (2010) Resveratrol and ascorbic acid prevent DNA damage induced by cryopreservation in human semen. Cryobiology 60:235–237PubMedCrossRefGoogle Scholar
  28. Brotherton J (1990) Cryopreservation of human semen. Arch Androl 25:181–195PubMedCrossRefGoogle Scholar
  29. Bucak MN, Uysal O (2008) The role of antioxidants in freezing of Saanen goat semen. Indian Vet J 85:148–150Google Scholar
  30. Camara DR, Mello-Pinto MMC, Pinto LC, Brasil OO, Nunes JF, Guerra MMP (2011) Effects of reduced glutathione and catalase on the kinematics and membrane functionality of sperm during liquid storage of ram semen. Small Rumin Res 100:44–49CrossRefGoogle Scholar
  31. Cerolini S, Maldjian A, Surai P, Noble R (2000) Viability, susceptibility to peroxidation and fatty acid composition of boar semen during liquid storage. Anim Reprod Sci 58:99–111PubMedCrossRefGoogle Scholar
  32. Chakrabarty J, Banerjee D, Pal D, De J, Ghosh A, Majumder GC (2007) Shedding off specific lipid constituents from sperm cell membrane during cryopreservation. Cryobiology 54:27–35PubMedCrossRefGoogle Scholar
  33. Chanapiwat P, Kaeoket K, Tummaruk P (2009) Effects of DHA-enriched hen egg yolk and l-cysteine supplementation on quality of cryopreserved boar semen. Asian J Androl 11:600–608PubMedPubMedCentralCrossRefGoogle Scholar
  34. Chatterjee S, Gagnon C (2001) Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol Reprod Dev 59:451–458PubMedCrossRefGoogle Scholar
  35. Chiachun T, Hong C, Haifun R (1991) The effects of selenium on gestation, fertility, and offspring in mice. Biol Trace Elem Res 30:227–231CrossRefGoogle Scholar
  36. Colas C, Junquera C, Perez-Pe R, Cebrian-Perez JA, Muino-Blanco T (2009) Ultrastructural study of the ability of seminal plasma proteins to protect ram spermatozoa against cold-shock. Microsc Res Tech 72:566–572PubMedCrossRefGoogle Scholar
  37. Çoyan K, Başpınar N, Bucak MN, Akalın PP (2011) Effects of cysteine and ergothioneine on post-thawed Merino ram sperm and biochemical parameters. Cryobiology 63:1–6PubMedCrossRefGoogle Scholar
  38. Çoyan K, Bucak MN, Başpınar N, Taşpınar M, Aydos S (2012) Ergothioneine attenuates the DNA damage of post-thawed Merino ram sperm. Small Rumin Res 106:165–167CrossRefGoogle Scholar
  39. Daghigh-Kia H, Olfati-Karaji R, Hoseinkhani A, Ashrafi I (2014) Effect of rosemary (Rosmarinus officinalis) extracts and glutathione antioxidants on bull semen quality after cryopreservation. Span J Agric Res 12:98–105CrossRefGoogle Scholar
  40. Dahl TA, Midden WR, Hartman PE (1988) Some prevalent biomolecules as defenses against singlet oxygen damage. Photochem Photobiol 47:357–362PubMedCrossRefGoogle Scholar
  41. de Paula TS, Bertolla RP, Spaine DM, Cunha MA, Schor N, Cedenho AP (2006) Effect of cryopreservation on sperm apoptotic deoxyribonucleic acid fragmentation in patients with oligozoospermia. Fertil Steril 86:597–600PubMedCrossRefGoogle Scholar
  42. Delmas D, Jannin B, Latruffe N (2005) Resveratrol: preventing properties against vascular alterations and ageing. Mol Nutr Food Res 49:377–395PubMedCrossRefGoogle Scholar
  43. Domínguez-Rebolledo A, Fernández-Santos M, García-Alvarez O, Maroto-Morales A, Garde J, Martínez-Pastor F (2009) Washing increases the susceptibility to exogenous oxidative stress in red deer spermatozoa. Theriogenology 72:1073–1084PubMedCrossRefGoogle Scholar
  44. Domínguez-Rebolledo ÁE et al (2010) Improving the effect of incubation and oxidative stress on thawed spermatozoa from red deer by using different antioxidant treatments. Reprod Fertil Dev 22:856–870PubMedCrossRefGoogle Scholar
  45. Donnelly ET, McClure N, Lewis SE (1999) Antioxidant supplementation in vitro does not improve human sperm motility. Fertil Steril 72:484–495PubMedCrossRefGoogle Scholar
  46. Dorostkar K, Alavi-Shoushtari SM, Mokarizadeh A (2012) Effects of in vitro selenium addition to the semen extender on the spermatozoa characteristics before and after freezing in water buffaloes (Bubalus bubalis). In: Veterinary Research Forum, vol 4. Faculty of Veterinary Medicine, Urmia University, Urmia, Iran, p 263Google Scholar
  47. Espino J, Bejarano I, Ortiz Á, Lozano GM, García JF, Pariente JA, Rodríguez AB (2010) Melatonin as a potential tool against oxidative damage and apoptosis in ejaculated human spermatozoa. Fertil Steril 94:1915–1917PubMedCrossRefGoogle Scholar
  48. Esteves SC, Sharma RK, Thomas AJ Jr, Agarwal A (2007) Evaluation of acrosomal status and sperm viability in fresh and cryopreserved specimens by the use of fluorescent peanut agglutinin lectin in conjunction with hypo-osmotic swelling test. Int Braz J Urol 33:364–376PubMedCrossRefGoogle Scholar
  49. Farshad A, Amidi F, Khor AK, Rashidi A (2011) Effect of Cholesterol-loaded-cyclodextrin in presence and absence of egg yolk during freezing step on quality of Markhoz Buck’s Spermatozoa. Asian-Australas J Anim Sci 24:181–189CrossRefGoogle Scholar
  50. Fernández-Santos MR et al (2007) Sperm characteristics and DNA integrity of Iberian red deer (Cervus elaphus hispanicus) epididymal spermatozoa frozen in the presence of enzymatic and nonenzymatic antioxidants. J Androl 28:294–305PubMedCrossRefGoogle Scholar
  51. Foote RH, Brockett CC, Kaproth MT (2002) Motility and fertility of bull sperm in whole milk extender containing antioxidants. Anim Reprod Sci 71:13–23PubMedCrossRefGoogle Scholar
  52. Frankel EN, Huang S-W, Aeschbach R, Prior E (1996) Antioxidant activity of a rosemary extract and its constituents, carnosic acid, carnosol, and rosmarinic acid, in bulk oil and oil-in-water emulsion. J Agric Food Chem 44:131–135CrossRefGoogle Scholar
  53. Fujinoki M (2008) Melatonin-enhanced hyperactivation of hamster sperm. Reproduction 136(5):533–541PubMedCrossRefGoogle Scholar
  54. Gadea J, Sellés E, Marco MA, Coy P, Matás C, Romar R, Ruiz S (2004) Decrease in glutathione content in boar sperm after cryopreservation: effect of the addition of reduced glutathione to the freezing and thawing extenders. Theriogenology 62:690–701PubMedCrossRefGoogle Scholar
  55. Gadea J, García-Vazquez F, Matás C, Gardón JC, Cánovas S, Gumbao D (2005) Cooling and freezing of boar spermatozoa: supplementation of the freezing media with reduced glutathione preserves sperm function. J Androl 26:396–404PubMedCrossRefGoogle Scholar
  56. Gadea J, Molla M, Selles E, Marco M, Garcia-Vazquez F, Gardon J (2011) Reduced glutathione content in human sperm is decreased after cryopreservation: effect of the addition of reduced glutathione to the freezing and thawing extenders. Cryobiology 62:40–46PubMedCrossRefGoogle Scholar
  57. Geva E, Bartoov B, Zabludovsky N, Lessing J, Lerner-Geva L, Amit A (1996) The effect of antioxidant treatment on human spermatozoa and fertilization rate in an in vitro fertilization program. Fertil Steril 66:430–434PubMedCrossRefGoogle Scholar
  58. Gil L et al (2010) Freezing ram semen: the effect of combination of soya and rosemary essences as a freezing extender on post-thaw sperm motility. Reprod Domest Anim 45:91CrossRefGoogle Scholar
  59. González N, Gil L, Martinez F, Malo C, Cano R, Mur P, Espinosa E (2010) Effect of natural antioxidant rosemary in canine soya freezing extender. Reprod Domest Anim 45:88Google Scholar
  60. Gwayi N, Bernard R (2002) The effects of melatonin on sperm motility in vitro in Wistar rats. Andrologia 34:391–396PubMedCrossRefGoogle Scholar
  61. Hammadeh M, Askari A, Georg T, Rosenbaum P, Schmidt W (1999) Effect of freeze-thawing procedure on chromatin stability, morphological alteration and membrane integrity of human spermatozoa in fertile and subfertile men. Int J Androl 22:155–162PubMedCrossRefGoogle Scholar
  62. Hatamoto LK, Sobrinho CB, Nichi M, Barnabe V, Barnabe R, Cortada C (2006) Effects of dexamethasone treatment (to mimic stress) and Vitamin E oral supplementation on the spermiogram and on seminal plasma spontaneous lipid peroxidation and antioxidant enzyme activities in dogs. Theriogenology 66:1610–1614PubMedCrossRefGoogle Scholar
  63. Horváthová E, Slameňová D, Navarová J (2010) Administration of rosemary essential oil enhances resistance of rat hepatocytes against DNA-damaging oxidative agents. Food Chem 123:151–156CrossRefGoogle Scholar
  64. Hu J-H, Tian W-Q, Zhao X-L, Zan L-S, Wang H, Li Q-W, Xin Y-P (2010) The cryoprotective effects of ascorbic acid supplementation on bovine semen quality. Anim Reprod Sci 121:72–77PubMedCrossRefGoogle Scholar
  65. Ip SP, Che CT, Leung PS (2001) Association of free radicals and the tissue renin-angiotensin system: prospective effects of Rhodiola, a genus of Chinese herb, on hypoxia-induced pancreatic injury. J Pancreas 2:16–25Google Scholar
  66. Isachenko V, Isachenko E, Katkov II, Montag M, Dessole S, Nawroth F, van der Ven H (2004) Cryoprotectant-free cryopreservation of human spermatozoa by vitrification and freezing in vapor: effect on motility, DNA integrity, and fertilization ability. Biol Reprod 71:1167–1173PubMedCrossRefGoogle Scholar
  67. Jang H et al (2010) Ameliorative effects of melatonin against hydrogen peroxide-induced oxidative stress on boar sperm characteristics and subsequent in vitro embryo development. Reprod Domest Anim 45:943–950PubMedCrossRefGoogle Scholar
  68. Jenkins TG, Aston KI, Carrell DT (2011) Supplementation of cryomedium with ascorbic acid–2-glucoside (AA2G) improves human sperm post-thaw motility. Fertil Steril 95:2001–2004PubMedCrossRefGoogle Scholar
  69. Jeong Y-J et al (2009) Effect of α-tocopherol supplementation during boar semen cryopreservation on sperm characteristics and expression of apoptosis related genes. Cryobiology 58:181–189PubMedCrossRefGoogle Scholar
  70. Johnson L, Weitze K, Fiser P, Maxwell W (2000) Storage of boar semen. Anim Reprod Sci 62:143–172PubMedCrossRefGoogle Scholar
  71. Kaeoket K, Chanapiwat P, Tummaruk P, Techakumphu M (2010) Supplemental effect of varying l-cysteine concentrations on the quality of cryopreserved boar semen. Asian J Androl 12:760PubMedPubMedCentralCrossRefGoogle Scholar
  72. Kalthur G, Raj S, Thiyagarajan A, Kumar S, Kumar P, Adiga SK (2011) Vitamin E supplementation in semen-freezing medium improves the motility and protects sperm from freeze-thaw–induced DNA damage. Fertil Steril 95:1149–1151PubMedCrossRefGoogle Scholar
  73. Kaya A, Aksoy M, Baspınar N, Yıldız C, Ataman M (2001) Effect of melatonin implantation to sperm donor rams on post-thaw viability and acrosomal integrity of sperm cells in the breeding and non-breeding season. Reprod Domest Anim 36:211–215PubMedCrossRefGoogle Scholar
  74. Kefer JC, Agarwal A, Sabanegh E (2009) Role of antioxidants in the treatment of male infertility. Int J Urol 16:449–457PubMedCrossRefGoogle Scholar
  75. Kirilova I, Ivanova-Kicheva M, Daskalova D, Gradinarska D, Kukov A, Dimitrov P, Hristova E (2015) The role of antioxidants and biologically active substances on the motility and speed parameters of buffalo bull spermatozoa after cryopreservation. Bulg J Agric Sci 21(1):209–214Google Scholar
  76. Kumar T, Kumar A, Swain DK, Yadav S (2011) Influence of oral supplementation of Zinc and Selenium on post thaw semen quality of Barbari bucks. J Anim Res 1(1)Google Scholar
  77. Kumi-Diaka J, Townsend J (2001) Effects of genistein isoflavone (4′,5′,7-trihydroxyisoflavone) and dexamethasone on functional characteristics of spermatozoa. J Med Food 4:39–47PubMedCrossRefGoogle Scholar
  78. Lasso JL, Noiles EE, Alvarez JG, Storey BT (1994) Mechanism of superoxide dismutase loss from human sperm cells during cryopreservation. J Androl 15:255PubMedGoogle Scholar
  79. Lewis S, Boyle P, McKinney K, Young I, Thompson W (1995) Total antioxidant capacity of seminal plasma is different in fertile and infertile men. Fertil Steril 64:868–870PubMedCrossRefGoogle Scholar
  80. Li P, Li Z-H, Dzyuba B, Hulak M, Rodina M, Linhart O (2010) Evaluating the impacts of osmotic and oxidative stress on common carp (Cyprinus carpio, L.) sperm caused by cryopreservation techniques. Biol Reprod 83:852–858PubMedCrossRefGoogle Scholar
  81. Linster CL, Van Schaftingen E (2007) Vitamin C. Biosynthesis, recycling and degradation in mammals. FEBS J 274(1):1–22PubMedCrossRefGoogle Scholar
  82. Liu Z, Zhang L, Ma H, Wang C, Li M, Wang Q (2005) Resveratrol reduces intracellular free calcium concentration in rat ventricular myocytes. Acta Physiol Sin Chin Edition 57:599Google Scholar
  83. Malo C, Gil L, Gonzalez N, Martínez F, Cano R, De Blas I, Espinosa E (2010) Anti-oxidant supplementation improves boar sperm characteristics and fertility after cryopreservation: comparison between cysteine and rosemary (Rosmarinus officinalis). Cryobiology 61:142–147PubMedCrossRefGoogle Scholar
  84. Malo C, Gil L, Cano R, Martínez F, Galé I (2011) Antioxidant effect of rosemary (Rosmarinus officinalis) on boar epididymal spermatozoa during cryopreservation. Theriogenology 75:1735–1741PubMedCrossRefGoogle Scholar
  85. Martínez-Páramo S, Diogo P, Dinis M, Herráez M, Sarasquete C, Cabrita E (2012) Incorporation of ascorbic acid and α-tocopherol to the extender media to enhance antioxidant system of cryopreserved sea bass sperm. Theriogenology 77:1129–1136PubMedCrossRefGoogle Scholar
  86. Martínez-Pastor F, Fernández-Santos M, Del Olmo E, Domínguez-Rebolledo A, Esteso M, Montoro V, Garde J (2008) Mitochondrial activity and forward scatter vary in necrotic, apoptotic and membrane-intact spermatozoan subpopulations. Reprod Fertil Dev 20:547–556PubMedCrossRefGoogle Scholar
  87. Martínez-Pastor F, Aisen E, Fernández-Santos MR, Esteso MC, Maroto-Morales A, García-Álvarez O, Garde JJ (2009) Reactive oxygen species generators affect quality parameters and apoptosis markers differently in red deer spermatozoa. Reproduction 137:225–235PubMedCrossRefGoogle Scholar
  88. Martín-Hidalgo D, Baron F, Bragado M, Carmona P, Robina A, Garcia-Marin L, Gil M (2011) The effect of melatonin on the quality of extended boar semen after long-term storage at 17 C. Theriogenology 75:1550–1560PubMedCrossRefGoogle Scholar
  89. Matsuoka T, Imai H, Kohno H, Fukui Y (2006) Effects of bovine serum albumin and trehalose in semen diluents for improvement of frozen-thawed ram spermatozoa. J Reprod Dev 52:675–683PubMedCrossRefGoogle Scholar
  90. Maxwell W, Stojanov T (1996) Liquid storage of ram semen in the absence or presence of some antioxidants. Reprod Fertil Dev 8:1013–1020PubMedCrossRefGoogle Scholar
  91. McNiven MA, Richardson GF (2006) Effect of quercetin on capacitation status and lipid peroxidation of stallion spermatozoa. Cell Preserv Technol 4:169–177CrossRefGoogle Scholar
  92. Mickle D, Weisel R (1992) Future directions of vitamin E and its analogues in minimizing myocardial ischemia-reperfusion injury. Can J Cardiol 9:89–93Google Scholar
  93. Minaei MB et al (2012) Effect of Trolox addition to cryopreservation media on human sperm motility. Iran J Reprod Med 10:99PubMedPubMedCentralGoogle Scholar
  94. Mook-Jung I, Kim H, Fan W, Tezuka Y, Kadota S, Nishijo H, Jung MW (2002) Neuroprotective effects of constituents of the oriental crude drugs, Rhodiola sacra, R. sachalinensis and Tokaku-joki-to, against beta-amyloid toxicity, oxidative stress and apoptosis. Biol Pharm Bull 25:1101–1104PubMedCrossRefGoogle Scholar
  95. Motohashi N, Mori I, Sugiura Y (1976) Complexing of copper ion by ergothioneine. Chem Pharm Bull 24:2364PubMedCrossRefGoogle Scholar
  96. Nagasaka R, Okamoto N, Ushio H (2004) Partial oxidative-stress perturbs membrane permeability and fluidity of fish nucleated red blood cells. Comp Biochem Physiol C Toxicol Pharmacol 139:259–266PubMedCrossRefGoogle Scholar
  97. Nekoonam S, Nashtaei MS, Zangi BM, Amidi F (2016) Effect of Trolox on sperm quality in normozospermia and oligozospermia during cryopreservation. Cryobiology 72:106–111PubMedCrossRefGoogle Scholar
  98. Niki E (1987) Interaction of ascorbate and alpha-tocopherol. Ann N Y Acad Sci 498:186–199PubMedCrossRefGoogle Scholar
  99. O’connell M, McClure N, Lewis S (2002) The effects of cryopreservation on sperm morphology, motility and mitochondrial function. Hum Reprod 17:704–709PubMedCrossRefGoogle Scholar
  100. Oehninger S, Duru NK, Srisombut C, Morshedi M (2000) Assessment of sperm cryodamage and strategies to improve outcome. Mol Cell Endocrinol 169:3–10PubMedCrossRefGoogle Scholar
  101. O’flaherty C, Beconi M, Beorlegui N (1997) Effect of natural antioxidants, superoxide dismutase and hydrogen peroxide on capacitation of frozen-thawed bull spermatozoa. Andrologia 29:269–275PubMedCrossRefGoogle Scholar
  102. Ohsugi M et al (1999) Active-oxygen scavenging activity of traditional nourishing-tonic herbal medicines and active constituents of Rhodiola sacra. J Ethnopharmacol 67:111–119PubMedCrossRefGoogle Scholar
  103. Ortiz A et al (2011) High endogenous melatonin concentrations enhance sperm quality and short-term in vitro exposure to melatonin improves aspects of sperm motility. J Pineal Res 50:132–139PubMedGoogle Scholar
  104. Panke ES, Rollag MD, Reitter RJ (1979) Pineal melatonin concentrations in the Syrian Hamster. Endocrinology 104:194–197PubMedCrossRefGoogle Scholar
  105. Partyka A, Łukaszewicz E, Niżański W (2012) Effect of cryopreservation on sperm parameters, lipid peroxidation and antioxidant enzymes activity in fowl semen. Theriogenology 77:1497–1504PubMedCrossRefGoogle Scholar
  106. Pena F, Johannisson A, Wallgren M, Martinez HR (2003) Antioxidant supplementation in vitro improves boar sperm motility and mitochondrial membrane potential after cryopreservation of different fractions of the ejaculate. Anim Reprod Sci 78:85–98PubMedCrossRefGoogle Scholar
  107. Ramadan T, Taha T, Samak M, Hassan A (2009) Effectiveness of exposure to longday followed by melatonin treatment on semen characteristics of Damascus male goats during breeding and non-breeding seasons. Theriogenology 71:458–468PubMedCrossRefGoogle Scholar
  108. Reiter RJ (1973) Pineal control of a seasonal reproductive rhythm in male golden hamsters exposed to natural daylight and temperature. Endocrinology 92:423–430PubMedCrossRefGoogle Scholar
  109. Roca J, Carvajal G, Lucas X, Vazquez JM, Martinez EA (2003) Fertility of weaned sows after deep intrauterine insemination with a reduced number of frozen-thawed spermatozoa. Theriogenology 60:77–87PubMedCrossRefGoogle Scholar
  110. Said TM, Gaglani A, Agarwal A (2010) Implication of apoptosis in sperm cryoinjury. Reprod Biomed Online 21:456–462PubMedCrossRefGoogle Scholar
  111. Sanders D (1984) Use of selenium in problem cattle herds. Mod Vet Pract 65:136–138PubMedGoogle Scholar
  112. Sarabia L, Maurer I, Bustos-Obregon E (2009) Melatonin prevents damage elicited by the organophosphorous pesticide diazinon on mouse sperm DNA. Ecotoxicol Environ Saf 72:663–668PubMedCrossRefGoogle Scholar
  113. Sharma RK, Agarwal A (1996) Role of reactive oxygen species in male infertility. Urology 48:835–850PubMedCrossRefGoogle Scholar
  114. Shiva M, Gautam AK, Verma Y, Shivgotra V, Doshi H, Kumar S (2011) Association between sperm quality, oxidative stress, and seminal antioxidant activity. Clin Biochem 44:319–324PubMedCrossRefGoogle Scholar
  115. Sierens J, Hartley J, Campbell M, Leathem A, Woodside J (2002) In vitro isoflavone supplementation reduces hydrogen peroxide-induced DNA damage in sperm. Teratog Carcinog Mutagen 22:227–234PubMedCrossRefGoogle Scholar
  116. Silva S, Soares A, Batista A, Almeida F, Nunes J, Peixoto C, Guerra M (2011) In vitro and in vivo evaluation of ram sperm frozen in tris egg-yolk and supplemented with superoxide dismutase and reduced glutathione. Reprod Domest Anim 46:874–881PubMedCrossRefGoogle Scholar
  117. Silva E, Cajueiro J, Silva S, Soares P, Guerra M (2012) Effect of antioxidants resveratrol and quercetin on in vitro evaluation of frozen ram sperm. Theriogenology 77:1722–1726PubMedCrossRefGoogle Scholar
  118. Sönmez M, Yüce A, Türk G (2007) The protective effects of melatonin and vitamin E on antioxidant enzyme activities and epididymal sperm characteristics of homocysteine treated male rats. Reprod Toxicol 23:226–231PubMedCrossRefGoogle Scholar
  119. Stojanović S, Sprinz H, Brede O (2001) Efficiency and mechanism of the antioxidant action of trans-resveratrol and its analogues in the radical liposome oxidation. Arch Biochem Biophys 391:79–89PubMedCrossRefGoogle Scholar
  120. Succu S, Berlinguer F, Pasciu V, Satta V, Leoni GG, Naitana S (2011) Melatonin protects ram spermatozoa from cryopreservation injuries in a dose-dependent manner. J Pineal Res 50:310–318PubMedCrossRefGoogle Scholar
  121. Szczęśniak-Fabiańczyk B, Bochenek M, Smorąg Z, Ryszka F (2003) Effect of antioxidants added to boar semen extender on the semen survival time and sperm chromatin structure. Reprod Biol 3:81–87PubMedGoogle Scholar
  122. Taylor K, Roberts P, Sanders K, Burton P (2009) Effect of antioxidant supplementation of cryopreservation medium on post-thaw integrity of human spermatozoa. Reprod Biomed Online 18:184–189PubMedCrossRefGoogle Scholar
  123. Therond P, Auger J, Legrand A, Jouannet P (1996) α-Tocopherol in human spermatozoa and seminal plasma: relationships with motility, antioxidant enzymes and leukocytes. Mol Hum Reprod 2:739–744PubMedCrossRefGoogle Scholar
  124. Thomson L, Fleming S, Aitken R, De Iuliis G, Zieschang J-A, Clark A (2009) Cryopreservation-induced human sperm DNA damage is predominantly mediated by oxidative stress rather than apoptosis. Hum Reprod dep214Google Scholar
  125. Uysal O, Bucak M (2007) Effects of oxidized glutathione, bovine serum albumin, cysteine and lycopene on the quality of frozen-thawed ram semen. Acta Vet Brno 76:383–390CrossRefGoogle Scholar
  126. Varghese A, Das S, Bhattacharya A, Bhattacharya S, Mandal M, Agarwal A (2005) Effect of cryoprotective additives-reduced glutathione, acetyl-L-carnitine on sperm membrane lipid peroxidation, DNA integrity and recovery of motile human sperm. Fertil Steril 84:S410–S411CrossRefGoogle Scholar
  127. Venkatesh S, Singh A, Shamsi MB, Thilagavathi J, Kumar R, Mitra DK, Dada R (2011) Clinical significance of sperm DNA damage threshold value in the assessment of male infertility. Reprod Sci 18:1005–1013PubMedCrossRefGoogle Scholar
  128. Wang AW, Zhang H, Ikemoto I, Anderson DJ, Loughlin KR (1997) Reactive oxygen species generation by seminal cells during cryopreservation. Urology 49:921–925PubMedCrossRefGoogle Scholar
  129. Weir CP, Robaire B (2007) Spermatozoa have decreased antioxidant enzymatic capacity and increased reactive oxygen species production during aging in the Brown Norway rat. J Androl 28:229–240PubMedCrossRefGoogle Scholar
  130. Wu T-W, Hashimoto N, Wu J, Carey D, Li R-K, Mickle DA, Weisel RD (1990) The cytoprotective effect of Trolox demonstrated with three types of human cells. Biochem Cell Biol 68:1189–1194PubMedCrossRefGoogle Scholar
  131. Yamamoto I, Muto N, Nagata E, Nakamura T, Suzuki Y (1990) Formation of a stable L-ascorbic acid α-glucoside by mammalian α-glucosidase-catalyzed transglucosylation. Biochim Biophys Acta (BBA) Gen Sub 1035:44–50CrossRefGoogle Scholar
  132. Yoshimoto T, Nakamura S, Yamauchi S, Muto N, Nakada T, Ashizawa K, Tatemoto H (2008) Improvement of the post-thaw qualities of Okinawan native pig spermatozoa frozen in an extender supplemented with ascorbic acid 2-O-α-glucoside. Cryobiology 57:30–36PubMedCrossRefGoogle Scholar
  133. Zeitoun MM, Al-Damegh MA (2014) Effect of Nonenzymatic Antioxidants on Sperm Motility and Survival Relative to Free Radicals and Antioxidant Enzymes of Chilled-Stored Ram Semen. Open J Anim Sci 5:50CrossRefGoogle Scholar
  134. Zhang J, Robinson D, Salmon P (2006) A novel function for selenium in biological system: selenite as a highly effective iron carrier for Chinese hamster ovary cell growth and monoclonal antibody production. Biotechnol Bioeng 95:1188–1197PubMedCrossRefGoogle Scholar
  135. Zhao H-w, Li Q-w, Ning G-z, Han Z-s, Jiang Z-l, Duan Y-f (2009) Rhodiola sacra aqueous extract (RSAE) improves biochemical and sperm characteristics in cryopreserved boar semen. Theriogenology 71:849–857PubMedCrossRefGoogle Scholar
  136. Zribi N, Chakroun NF, El Euch H, Gargouri J, Bahloul A, Keskes LA (2010) Effects of cryopreservation on human sperm deoxyribonucleic acid integrity. Fertil Steril 93:159–166PubMedCrossRefGoogle Scholar
  137. Zribi N et al (2012) Effect of freezing–thawing process and quercetin on human sperm survival and DNA integrity. Cryobiology 65:326–331PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Fardin Amidi
    • 1
  • Azar Pazhohan
    • 1
  • Maryam Shabani Nashtaei
    • 1
    Email author
  • Mahshad Khodarahmian
    • 1
  • Saeid Nekoonam
    • 1
  1. 1.Department of Anatomy, School of MedicineTehran University of Medical SciencesTehranIran

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